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Rising costs and a depleting supply of oil, as well as environmental concerns, have led to strong interest in renewable fuels and chemicals. In the last decade, there was considerable interest for microbial production of biofuels by metabolic engineering approach as an attractive alternative to transportation fuels. Recent advancements of synthetic biology have led to the further development of genetic engineering of microorganism, which has been great motivation for developing strategy for microbial production of biofuels. Rational design of metabolism is very important for production of biofuels.